Simplified 25-Hydroxyvitamin D Standardization and Optimization in Dried Blood Spots by LC-MS/MS

2017 ◽  
Vol 100 (5) ◽  
pp. 1328-1336 ◽  
Author(s):  
Andrew J Makowski ◽  
John A Rathmacher ◽  
Ronald L Horst ◽  
Christopher T Sempos

Abstract Previous studies have assessed vitamin D status based on the 25-hydroxyvitamin D [25(OH)D] concentration measured in samples from dried blood spots (DBSs). In 40 individuals participating in a clinical study, we compared 25(OH)D levels measured from DBSs and in serum using an LC-MS/MS reference procedure in collaboration with the Vitamin D Standardization Program. The main objective was to simplify and optimize current methods to produce an assay that can be used as a screening tool for 25(OH)D concentration assessment without derivatization. The DBS 25(OH)D levels, compared to serum concentrations, were found to have 101% accuracy overall, and the correlation coefficient (r) was 0.83 (P < 0.0001), with a significant linear relationship. Free 25(OH)D and vitamin D binding protein (VDBP) were assessed in the serum samples for potential correlations to the DBS calculations: the levels of free 25(OH)D had moderate to strong correlation to DBS and serum concentrations, with r values of 0.67 (P < 0.0001) and 0.76 (P < 0.0001), respectively. VDBP and hematocrit had no significant correlation to either DBS or serum sample types, with r values <0.1. In conclusion, the useof two DBSs and an increase in DBS sample size improved overall sample representation without the need for derivatization, and produced an accurate and robust method that can be used to screen 25(OH)D levels.

2016 ◽  
Vol 5 ◽  
Author(s):  
Lauren R. Young ◽  
Robert C. Backus

AbstractLittle is known regarding optimal vitamin D status in adult dogs. To date no studies on vitamin D supplementation for improving vitamin D status have been reported for adult dogs. The aims of this study were to identify dogs with low vitamin D status and evaluate an oral dosage of cholecalciferol (D3) for effectiveness in increasing vitamin D status. For this, forty-six privately owned dogs were evaluated. Of the dogs, thirty-three (or 71·7 %) had serum 25-hydroxyvitamin D (25(OH)D) concentrations less than 100 ng/ml, a minimum previously suggested for vitamin D sufficiency in dogs. Subsequently, thirteen dogs were enrolled in a supplementation trial. Dogs were given either a D3supplement (n7; 2·3 µg/kg0·75) or olive oil placebo (n6) daily with food. Serum concentrations of 25(OH)D were determined at weeks 1, 3 and 6, and at the trial end. Only at the trial end (weeks 9–10) was 25(OH)D significantly greater (P= 0·05) in supplementedv.placebo dogs. Serum concentrations of 24R,25-dihydroxycholecalciferol determined at the trial end were about 40 % of that of 25(OH)D3and not significantly different between the groups. Concentrations of parathyroid hormone, ionised Ca, P and creatinine measured in initial and final serum samples indicated supplementation caused no toxicity. We conclude that vitamin D3supplementation at a dosage near the National Research Council recommended safe-upper limit was not effective for rapidly raising serum 25(OH)D concentrations in healthy, adult dogs. Further work is needed in evaluating the metabolism of orally administered D3in dogs before dosing recommendations can be made.


2016 ◽  
Vol 62 (4) ◽  
pp. 639-646 ◽  
Author(s):  
David Kvaskoff ◽  
Alicia K Heath ◽  
Henry A Simila ◽  
Pauline Ko ◽  
Dallas R English ◽  
...  

Abstract BACKGROUND The noncalcemic actions of vitamin D in multiple organs are now widely recognized. Vitamin D status has been linked with a wide variety of conditions, which has led to an increasing demand for vitamin D screening. In particular, there is intense interest in the impact of vitamin D on a variety of developmental conditions. The most readily accessible pediatric samples are dried blood spots, and health organizations are increasingly archiving such samples for later assessment of the antecedents of disease. METHODS In 2009, we developed a method to quantify the major circulatory form of vitamin D, 25-hydroxyvitamin D, in archived dried blood spots. Over the last 6 years, we have made substantial alterations to the published method to enhance throughput, sensitivity, and assay robustness. RESULTS With the alterations, the assay was 3 times faster than the previously published assay and had a >10-fold increase in signal strength. Intraassay imprecision decreased from 13.4% to 6.9%, and there was a 5-fold reduction in interfering phospholipids. In actual use over 2 years, the assay showed an interassay imprecision of 11.6%. CONCLUSIONS This assay has performed reliably over the past 6 years. The practical changes we have made should allow clinical chemists to successfully adapt this method.


2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Hou-Qun Ying ◽  
Hui-Ling Sun ◽  
Bang-Shun He ◽  
Yu-Qin Pan ◽  
Feng Wang ◽  
...  

Nutrients ◽  
2021 ◽  
Vol 13 (5) ◽  
pp. 1714
Author(s):  
Habiba AlSafar ◽  
William B. Grant ◽  
Rafiq Hijazi ◽  
Maimunah Uddin ◽  
Nawal Alkaabi ◽  
...  

Insufficient blood levels of the neurohormone vitamin D are associated with increased risk of COVID-19 severity and mortality. Despite the global rollout of vaccinations and promising preliminary results, the focus remains on additional preventive measures to manage COVID-19. Results conflict on vitamin D’s plausible role in preventing and treating COVID-19. We examined the relation between vitamin D status and COVID-19 severity and mortality among the multiethnic population of the United Arab Emirates. Our observational study used data for 522 participants who tested positive for SARS-CoV-2 at one of the main hospitals in Abu Dhabi and Dubai. Only 464 of those patients were included for data analysis. Demographic and clinical data were retrospectively analyzed. Serum samples immediately drawn at the first hospital visit were used to measure serum 25-hydroxyvitamin D [25(OH)D] concentrations through automated electrochemiluminescence. Levels < 12 ng/mL were significantly associated with higher risk of severe COVID-19 infection and of death. Age was the only other independent risk factor, whereas comorbidities and smoking did not contribute to the outcomes upon adjustment. Sex of patients was not an important predictor for severity or death. Our study is the first conducted in the UAE to measure 25(OH)D levels in SARS-CoV-2-positive patients and confirm the association of levels < 12 ng/mL with COVID-19 severity and mortality.


2018 ◽  
Vol 144 (10) ◽  
pp. 2401-2407 ◽  
Author(s):  
Chen Yuan ◽  
Irene M. Shui ◽  
Kathryn M. Wilson ◽  
Meir J. Stampfer ◽  
Lorelei A. Mucci ◽  
...  

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